Germ Cell Differentiation from Human Embryonic Stem Cells

大约10%-15%的夫妇患有不孕症，其原因与本地区所有重大事件有关 生殖途径，从发育有功能的卵子和精子到受精，胚胎 转录激活、着床、胎盘形成和存活胎儿的生长。尽管频率很高 不孕不育及其对患者生活质量的影响，对潜在的分子和遗传因素知之甚少 原因。无可争议，辅助生殖技术、生殖生物学、人类 胚胎干细胞(HESC)生物学和人类遗传学，现在可能允许我们克服两个具有历史意义的 人类生殖研究的局限性：即无法获得早期人类发育 到生物探索和人类基因组在发育过程中的遗传难题。在这 建议，我们使用hESC生物学和人类遗传学的强大工具来检验假设：1) DAZ(在无精子症中缺失)基因家族需要多个成员才能形成和/或 维持新生的人类生殖细胞和2)常见的遗传变异和独特的突变， 富含不育男性和女性的染色体，调节双方早期人类生殖细胞的发育 性别。我们的具体目标是：目标1.确定基线分子、时间和遗传特征 体内外hESCs和mESCs向生殖细胞分化的研究目标2.让人类达兹安静下来， 以及人类和小鼠的DAZL和BOL基因，并评估生殖细胞发育。目标3.检查功能 DAZ基因家族常见变异和罕见突变在人群中的意义 将结果与人类基因研究和临床发现联系起来。这项研究尤其对 寻求辅助生殖技术的不孕夫妇的健康担忧 亲生父母的身份。事实上，成功完成拟议的研究有望加强我们的 对人类生殖细胞发育的重要途径的基本了解，提供了有用的工具 为基础科学家研究人类生殖细胞发育，并为小说的发展做出贡献 临床使用的治疗学和有效的诊断基因测试。这项研究将大大受益于 建立一个中心，提供整个社区的互动并访问有价值的核心 设施。本研究使用美国国立卫生研究院注册中心的hESC品系(UC01、UC06、WA01和WA09)。

Infertility afflicts approximately 10-15% of couples and has causes linked to all the major events in the reproductive pathways, from development of functional eggs and sperm, to fertilization, embryonic transcriptional activation, implantation, placentation and growth of a viable fetus. In spite of the frequency of infertility and impact on the quality of life of those affected, little is known of underlying molecular and genetic causes. Nonethless, recent advances in assisted reproductive techniques, reproductive biology, human embryonic stem cell (hESC) biology and human genetics, may now allow us to overcome two historicallysignificant limitations in human reproductive studies: namely, the inaccessibility of early human development to biological exploration and the genetic-intractability of the human genome during development. In this proposal, we use the powerful tools of hESC biology and human genetics to test the hypotheses: 1) that multiple members of the DAZ (Deleted in AZoospermia) gene family are required for the formation and/or maintenance of nascent human germ cells and 2) that common genetic variants and unique mutations, enriched in chromosomes of infertile men and women, modulate early human germ cell development in both sexes. Our specific aims are: Aim 1. Define the baseline molecular, temporal and genetic characteristics of differentiation of hESCs and mESCs to the germ cell lineage in vitro and in.vivo. Aim 2. Silence human DAZ, and human and mouse DAZL and BOL genes and assess germ cell development. Aim 3. Examine functional significance of a common variant and a rare mutation in the DAZ gene family in the human population and relate results to human genetic studies and clinical findings. This research is particularly responsive to the health concerns of infertile couples who seek assisted reproductive technologies in hopes of achieving biological parenthood. Indeed, successful completion of the proposed research promises to strengthen our basic understanding of the remarkable pathways by which human germ cells develop, provide useful tools for basic scientists to study human germ cell development, and contribute to the development of novel therapeutics and validated diagnostic genetic tests for clinical use. The research would greatly benefit from the establishment of a center that provides interactions across the community and access to valuable core facilities. This research uses hESC lines from the NIH Registry (UC01, UC06, WA01 and WA09).